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Handmade leaf cutter for efficient and reliable ROS assay.

Yoshitake Desaki^1,2, Hikaru Shimada¹, Shohei Takahashi¹

¹Department of Life Sciences, School of Agriculture, Meiji University, 1-1-1 Higashi-Mita, Tama-ku, Kawasaki, Kanagawa, 214-8571, Japan.

Plant Biotechnology (Tokyo, Japan)

|January 28, 2020

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This study introduces a handmade leaf cutter to improve the reliability of reactive oxygen species (ROS) analysis in plant immunity. The new method enhances reproducibility for MAMP/elicitor-induced ROS assays across various plant species.

Area of Science:

Plant biology
Plant immunity
Biochemical assays

Background:

Reactive oxygen species (ROS) generation is a key indicator of plant immune responses.
The leaf disk assay is widely used for MAMP/elicitor-induced ROS analysis but suffers from reproducibility issues.
Current methods are challenging for plants with tough leaves due to poor MAMP/elicitor penetration.

Purpose of the Study:

To develop a more reliable and reproducible method for MAMP/elicitor-induced ROS analysis.
To overcome limitations of the traditional leaf disk assay, especially for recalcitrant plant species.
To introduce a novel tool for standardized plant immune response studies.

Main Methods:

Development of a handmade leaf cutter to create uniform leaf fragments with slits.

Keywords:

MAMP ROS defense response elicitor leaf disk

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Application of the leaf cutter in MAMP/elicitor-induced ROS assays.

Testing the method's efficacy on various plant species, including rice.

Main Results:

The handmade leaf cutter produced uniform leaf fragments, improving assay reliability.
Increased numbers of leaf fragments further enhanced the reproducibility of ROS detection.
The method demonstrated successful application to rice, indicating broad plant species applicability.

Conclusions:

The novel leaf cutter significantly improves the reliability and reproducibility of the leaf disk assay for ROS analysis.
This technique is suitable for a wider range of plant species, including those with tough leaf structures.
The improved assay facilitates more accurate assessment of plant immune responses to MAMPs/elicitors.